Diphenylazetidinone with improved physiological properties, process for its preparation, medicaments comprising this compound, and its use

ABSTRACT

The invention relates to the compound of the formula 1 
                         
and to its physiologically acceptable salts. The compound is suitable, for example, as a hypolipidemic.

This application is a continuation of application Ser. No. 10/813,954,filed Mar. 31, 2004, now U.S. Pat. No. 7,205,290.

The invention relates to a substituted diphenylazetidinone, to itsphysiologically acceptable salts and to physiologically functionalderivatives.

Diphenylazetidinones and their use for treating hyperlipidemia and alsoarteriosclerosis and hypercholesterolemia have already been described(WO 02/50027).

It was an object of the invention to provide a compound which, comparedto the compounds described in WO 02/50027, has considerably bettersolubility in the upper small intestine in the pre- and/or postprandialstate. The improved solubility of the compound ensures higheravailability of dissolved substance at the site of action and thusimproved efficacy.

FaSSIF (fasted State Simulating Intestinal Fluid) and FeSSIF (Fed StateSimulating Intestinal Fluid) media which reflect the pH/solubilizationconditions in the upper small intestine in the pre- and postprandialstate, respectively, were used to test this improved solubility.

It was another object of the invention to provide a compound which,compared to the compounds described in WO 02/50027, has increasedstability both in the acidic range (stomach) and in the weakly alkalinerange (small intestine). This property leads to fewerbyproducts/cleavage products which for their part may have unwantedside-effects. However, increased stability in the acidic range is also agreat advantage for formulation since in this case there is no need foran acid-resistant capsule/tablet.

Accordingly, the invention relates to the compounds of the formula I

and pharmaceutically acceptable salts thereof.

Because they are more soluble in water than the starting compounds orbasis compounds, pharmaceutically acceptable salts are particularlysuitable for medical applications. These salts must possess apharmaceutically acceptable anion or cation. Suitable pharmaceuticallyacceptable acid addition salts of the compound according to theinvention are salts of inorganic acids, such as hydrochloric acid,hydrobromic acid, phosphoric acid, metaphosphoric acid, nitric acid andsulfuric acid, and also of organic acids, such as acetic acid,benzenesulfonic acid, benzoic acid, citric acid, ethanesulfonic acid,fumaric acid, gluconic acid, glycolic acid, isethionic acid, lacticacid, lactobionic acid, maleic acid, malic acid, methanesulfonic acid,succinic acid, p-toluenesulfonic acid and tartaric acid. Suitablepharmaceutically acceptable basic salts are ammonium salts, alkali metalsalts (such as sodium salts and potassium salts), alkaline earth metalsalts (such as magnesium salts and calcium salts), trometamol(2-amino-2-hydroxymethyl-1,3-propanediol), diethanolamine, lysine orethylenediamine.

Salts containing an anion which is not pharmaceutically acceptable, suchas, for example, trifluoroacetate, also belong within the scope of theinvention as useful intermediates for preparing or purifyingpharmaceutically acceptable salts and/or for use in nontherapeuticapplications, for example in-vitro applications.

As used herein, the following definitions apply:

“Patient” means a warm blooded animal, such as for example rat, mice,dogs, cats, guinea pigs, and primates such as humans.

“Treat” or “treating” means to alleviate symptoms, eliminate thecausation of the symptoms either on a temporary or permanent basis, orto prevent or slow the appearance of symptoms of the named disorder orcondition.

“Therapeutically effective amount” means a quantity of the compoundwhich is effective in treating the named disorder or condition.

“Pharmaceutically acceptable carrier” is a non-toxic solvent,dispersant, excipient, adjuvant or other material which is mixed withthe active ingredient in order to permit the formation of apharmaceutical composition, i.e., a dosage form capable ofadministration to the patient. One example of such a carrier is apharmaceutically acceptable oil typically used for parenteraladministration.

The term “physiologically functional derivative”, denotes anyphysiologically acceptable derivative of a compound of the formula Iaccording to the invention, e.g. an ester which is able, on beingadministered to a mammal, such as a human, to form (directly orindirectly) a compound of the formula I or an active metabolite thereof.

The physiologically functional derivatives also include prodrugs of thecompound according to the invention, as described, for example, in H.Okada et al., Chem. Pharm. Bull. 1994, 42, 57-61. Such prodrugs can bemetabolized in vivo to give a compound according to the invention. Theseprodrugs may or may not themselves be active.

The compound according to the invention can also be present in differentpolymorphic forms, for example as amorphous and crystalline polymorphicforms. All the polymorphic forms of the compound according to theinvention belong within the scope of the invention and are anotheraspect of the invention.

In that which follows, all references to “compound(s) according toformula I” relate to (a) compound(s) of the formula I as described aboveand to its (their) salts, solvates and physiologically functionalderivatives as described herein.

An aryl radical is to be understood as meaning a phenyl, naphthyl,biphenyl, tetrahydronaphthyl, alpha- or beta-tetralone, indanyl orindan-1-onyl radical.

The compound(s) of the formula (I) can also be administered incombination with (an) other active compound(s).

The quantity of a compound according to formula I which is required inorder to achieve the desired biological effect depends on a number offactors, e.g. the specific compound which is selected, the intended use,the type of administration and the clinical state of the patient. Ingeneral, the daily dose lies in a range from 0.01 mg to 100 mg(typically from 0.05 mg to 50 mg) per day per kilogram of body weight,e.g. 0.1-10 mg/kg/day.

Single dose formulations which can be administered orally, such astablets or capsules, can, for example, contain from 1.0 to 1000 mg,typically from 10 to 600 mg. While the compounds according to formula Ican themselves be used as the compound for treating the abovementionedconditions, they are preferably present, together with an acceptablecarrier, in the form of a pharmaceutical composition. The carriernaturally has to be acceptable, in the sense that it is compatible withthe other constituents of the composition and is not harmful to thehealth of the patient. The carrier can be a solid or a liquid or bothand is preferably formulated with the compound as a single dose, forexample as a tablet which can contain from 0.05% to 95% by weight of theactive compound. Other pharmaceutically active substances can also bepresent, including other compounds according to formula I. Thepharmaceutical compositions according to the invention can be preparedusing one of the known pharmaceutical methods, which essentially consistin mixing the constituents with pharmacologically acceptable carriersubstances and/or auxiliary substances.

Pharmaceutical compositions according to the invention are those whichare suitable for oral and peroral (e.g. sublingual) administration evenif the most suitable mode of administration depends, in each individualcase, on the nature and severity of the condition to be treated and onthe nature of the compound according to formula I which is in each caseemployed. Sugar-coated formulations and sugar-coated delayed-releaseformulations also belong within the scope of the invention. Formulationswhich are acid-resistant and gastric juice-resistant are preferred.Suitable gastric juice-resistant coatings include cellulose acetatephthalate, polyvinyl acetate phthalate, hydroxypropylmethyl cellulosephthalate and anionic polymers of methacrylic acid and methylmethacrylate.

Suitable pharmaceutical compounds for oral administration can be presentin separate units, such as capsules, cachets, lozenges or tablets whichin each case contain a specific quantity of the compound according toformula I; as powders or granulates; as a solution or suspension in anaqueous or non-aqueous liquid; or as an oil-in-water or a water-in-oilemulsion. As has already been mentioned, these compositions can beprepared using any suitable pharmaceutical method which includes a stepin which the active compound and the carrier (which can consist of oneor more additional constituents) are brought into contact. In general,the compositions are prepared by uniformly and homogeneously mixing theactive compound with a liquid and/or finely divided solid carrier, afterwhich the product is molded, if necessary. Thus, a tablet can beprepared, for example, by means of a powder or granulate of the compoundbeing pressed or molded, where appropriate together with one or moreadditional constituents. Pressed tablets can be prepared by tabletingthe compound in freely flowing form, such as a powder or granulate,which is mixed, where appropriate, with a binder, lubricant, inertdiluent and/or a (several) surface-active/dispersing agent(s) in asuitable machine. Molded tablets can be prepared by molding thepulverulent compound, which is moistened with an inert, liquid diluent,in a suitable machine.

Pharmaceutical compositions which are suitable for peroral (sublingual)administration include lozenges which contain a compound according toformula I together with a flavoring agent, usually sucrose and gumarabic or tragacanth, and pastils, which include the compound in aninert base such as gelatin and glycerol or sucrose and gum arabic.

The following are suitable to use as additional active compounds for thecombination preparations:

all the antidiabetics which are named in chapter 12 in the Roten Liste[Red List] 2003. They can be combined with compounds of the formula Iaccording to the invention, particularly for the purpose ofsynergistically improving the effect. The active compound combinationcan be administered either by separately administering the activecompounds to the patient or administering them in the form ofcombination preparations in which several active compounds are presentin one pharmaceutical preparation. Most of the active compounds whichare cited below are disclosed in USP Dictionary of USAN andInternational Drug Names, US Pharmacopeia, Rockville 2001.

Antidiabetics include insulin and insulin derivatives, such as Lantus®(see www.lantus.com) or HMR 1964, rapidly acting insulins (see U.S. Pat.No. 6,221,633), GLP-1 derivatives, such as those which Novo Nordisk A/Shas disclosed in WO 98/08871, Zealand has disclosed in WO/04156 andBeaufour-Ipsen has disclosed in WO 00/34331 and also orally activehypoglycaemic active compounds.

The orally active hypoglycaemic active compounds preferably includesulfonylureas, biguanidines, meglitinides, oxadiazolidinediones,thiazolidinediones, glucosidase and glycogen phosphorylase inhibitors,glucagon antagonists, GLP-1-agonists, potassium channel openers, such asthose which Novo Nordisk A/S has disclosed in WO 97/26265 and WO99/03861, insulin sensitizers, inhibitors of liver enzymes which areinvolved in stimulating gluconeogenesis and/or glycogenolysis,modulators of glucose uptake, glucose transport and glucosereabsorption, compounds which alter fat metabolism, such asantihyperlipidaemic active compounds and antilipidaemic activecompounds, compounds which decrease food intake, PPAR agonists and PXRagonists, and active compounds which act on the ATP-dependent potassiumchannel in the beta cells.

In one embodiment of the invention, the compounds of the formula I areadministered in combination with an HMGCoA reductase inhibitor, such assimvastatin, fluvastatin, pravastatin, lovastatin, atorvastatin,cerivastatin or rosuvastatin.

In one embodiment of the invention, the compounds of the formula I areadministered in combination with a cholesterol absorption inhibitor,such as ezetimibe, tiqueside or pamaqueside.

In one embodiment of the invention, the compounds of the formula I areadministered in combination with a PPAR gamma agonist, such asrosiglitazone, pioglitazone, JTT-501 or GI 262570.

In one embodiment of the invention, the compounds of the formula I areadministered in combination with a PPAR alpha agonist, such as GW 9578or GW 7647.

In one embodiment of the invention, the compounds of the formula I areadministered in combination with a mixed PPAR alpha/gamma agonist, suchas GW 1536, AVE 8042, AVE 8134 or AVE 0847, or as described inPCT/US/11833, PCT/US/11490 or DE10142734.4.

In one embodiment of the invention, the compounds of the formula I areadministered in combination with a fibrate, such as fenofibrate,clofibrate or bezafibrate.

In one embodiment of the invention, the compounds of the formula I areadministered in combination with an MTP inhibitor, such as implitapide,BMS-201038 or R-103757.

In one embodiment of the invention, the compounds of the formula I areadministered in combination with a bile acid absorption inhibitor (see,e.g., U.S. Pat. No. 6,245,744 or U.S. Pat. No. 6,221,897), such as HMR1741.

In one embodiment of the invention, the compounds of the formula I areadministered in combination with a CETP inhibitor, such as JTT-705.

In one embodiment of the invention, the compounds of the formula I areadministered in combination with a polymeric bile acid adsorber, such ascholestyramine or colesevelam.

In one embodiment of the invention, the compounds of the formula I areadministered in combination with an LDL receptor inducer (see U.S. Pat.No. 6,342,512), such as HMR1171 or HMR 1586.

In one embodiment of the invention, the compounds of the formula I areadministered in combination with an ACAT inhibitor, such as avasimibe.

In one embodiment of the invention, the compounds of the formula I areadministered in combination with an antioxidant, such as OPC-14117.

In one embodiment of the invention, the compounds of the formula I areadministered in combination with a lipoprotein lipase inhibitor, such asNO-1886

In one embodiment of the invention, the compounds of the formula I areadministered in combination with an ATP citrate lyase inhibitor, such asSB-204990.

In one embodiment of the invention, the compounds of the formula I areadministered in combination with a squalene synthetase inhibitor, suchas BMS-188494.

In one embodiment of the invention, the compounds of the formula I areadministered in combination with a lipoprotein(a) antagonist, such asCI-1027 or nicotinic acid.

In one embodiment of the invention, the compounds of the formula I areadministered in combination with a lipase inhibitor, such as orlistat.

In one embodiment of the invention, the compounds of the formula I areadministered in combination with insulin.

In one embodiment, the compounds of the formula I are administered incombination with a sulfonylurea, such as tolbutamide, glibenclamide,glipizide or glimepiride.

In one embodiment, the compounds of the formula I are administered incombination with a biguanide, such as metformin.

In yet another embodiment, the compounds of the formula I areadministered in combination with a meglitinide, such as repaglinide.

In one embodiment, the compounds of the formula I are administered incombination with a thiazolidinedione, such as troglitazone, ciglitazone,pioglitazone, rosiglitazone or the compounds disclosed by Dr. Reddy'sResearch Foundation in WO 97/41097, in particular5-[[4-[(3,4-dihydro-3-methyl-4-oxo-2-quinazolinylmethoxy]phenyl]methyl]-2,4-thiazolidinedione.

In one embodiment, the compounds of the formula I are administered incombination with an α-glucosidase inhibitor, such as miglitol oracarbose.

In one embodiment, the compounds of the formula I are administered incombination with an active compound which acts on the ATP-dependentpotassium channel of beta cells, such as tolbutamide, glibenclamide,glipizide, glimepiride or repaglinide.

In one embodiment, the compounds of the formula I are administered incombination with more than one of the abovementioned compounds, e.g. incombination with a sulfonylurea and metformin, a sulfonylurea andacarbose, repaglinide and metformin, insulin and a sulfonylurea, insulinand metformin, insulin and troglitazone, insulin and lovastatin, etc.

In another embodiment, the compounds of the formula I are administeredin combination with CART modulators (see “cocaine-amphetamine-regulatedtranscript influences energy metabolism, anxiety and gastric emptying inmice” Asakawa, A, et al., M.: Hormone and Metabolic Research (2001),33(9), 554-558), NPY antagonists, e.g. naphthalene-1-sulfonicacid-{4-[(4-aminoquinazolin-2-ylamino)methyl]cyclohexylmethyl}amidehydrochloride (CGP 71683A)), cannabinoid receptor 1 antagonists (see,e.g., EP 0656354, WO 00/15609 or WO 02/076949) MC4 agonists (e.g.1-amino-1,2,3,4-tetrahydronaphthalene-2-carboxylic acid[2-(3a-benzyl-2-methyl-3-oxo-2,3,3a,4,6,7-hexahydropyrazolo[4,3-c]pyridin-5-yl)-1-(4-chlorophenyl)-2-oxoethyl]amide;(WO 01/91752)), orexin antagonists (e.g.1-(2-methylbenzoxazol-6-yl)-3-[1,5]naphthyridin-4-yl urea hydrochloride(SB-334867-A)), H3 agonists(3-cyclohexyl-1-(4,4-dimethyl-1,4,6,7-tetrahydroimidazo[4,5-c]pyridin-5-yl)-propan-1-oneoxalic acid salt (WO 00/63208)); TNF agonists, CRF antagonists (e.g.[2-methyl-9-(2,4,6-trimethylphenyl)-9H-1,3,9-triazafluoren-4-yl]dipropylamine(WO 00/66585)), CRF BP antagonists (e.g. urocortin), urocortin agonists,β3-agonists (e.g.1-(4-chloro-3-methanesulfonylmethylphenyl)-2-[2-(2,3-dimethyl-1H-indol-6-yloxy)ethylamino]ethanolhydrochloride (WO 01/83451)), MSH (melanocyte-stimulating hormone)agonists, MCH (melanin-concentrating hormone) receptor antagonists (see,e.g., WO 03/15769), CCK-A agonists (e.g.{2-[4-(4-chloro-2,5-dimethoxyphenyl)-5-(2-cyclohexylethyl)thiazol-2-ylcarbamoyl]-5,7-dimethylindol-1-yl}aceticacid trifluoroacetic acid salt (WO 99/15525), or SR-146131 (WO 0244150)or SSR-125180), serotonin reuptake inhibitors (e.g. dexfenfluramine),mixed serotoninergic and noradrenergic compounds (e.g. WO 00/71549), 5HTagonists, e.g. 1-(3-ethylbenzofuran-7-yl)piperazine oxalic acid salt (WO01/09111), bombesin agonists, galanin antagonists, growth hormone (e.g.human growth hormone), growth hormone-releasing compounds (tert-butyl6-benzyloxy-1-(2-diisopropylaminoethylcarbamoyl)-3,4-dihydro-1H-isoquinoline-2-carboxylate(WO 01/85695)), TRH agonists (see, e.g. EP 0 462 884) uncoupling protein2 or protein 3 modulators, leptin agonists (see, e.g. Lee, Daniel W.;Leinung, Matthew C.; Rozhavskaya-Arena, Marina; Grasso, Patricia. Leptinagonists as a potential approach to the treatment of obesity. Drugs ofthe Future (2001), 26(9), 873-881), DA agonists (bromocriptine,doprexin), lipase/amylase inhibitors (erg. WO 00/40569), PPAR modulators(e.g. WO 00/78312), 11β-HSD1 (11-beta-hydroxysteroiddehydrogenasetype 1) inhibitors (see e.g. WO 01/90094 or T. Barf et al., J. Med.Chem. (2002), 45, 3813-3815), acetyl-CoA carboxylase (ACC; see e.g. WO99/46262) inhibitors, dipeptidylpeptidase IV (DPP-IV; see e.g. EP1259246) inhibitors, RXR modulators or TR-β-agonists.

In one embodiment of the invention, the other active compound is leptin;see, e.g., “Perspectives in the therapeutic use of leptin”, Salvador,Javier; Gomez-Ambrosi, Javier; Fruhbeck, Gema, Expert Opinion onPharmacotherapy (2001), 2(10), 1615-1622.

In one embodiment, the other active compound is dexamphetamine oramphetamine.

In one embodiment, the other active compound is fenfluramine ordexfenfluramine.

In yet another embodiment, the other active compound is sibutramine.

In one embodiment, the other active compound is orlistat.

In one embodiment, the other active compound is mazindol or phentermine.

In one embodiment, the compounds of the formula I are administered incombination with ballast substances, preferably insoluble ballastsubstances (see, e.g., carob/Caromax®) (Zunft H J; et al., Carob pulppreparation for treatment of hypercholesterolemia, ADVANCES IN THERAPY(2001 September-October), 18(5), 230-6). Caromax is a carob-containingproduct from Nutrinova, Nutrition Specialties & Food Ingredients GmbH,Industriepark Hoechst, 65926 Frankfurt/Main)). The combination withCaromax® can be effected in one preparation or by means of separatingadministering compounds of the formula I and Caromax®. In thisconnection, Caromax® can also be administered in the form of foodstuffs,for example in bread, cakes and pastries or muesli bars.

It will be understood that each suitable combination of the compoundsaccording to the invention with one or more of the abovementionedcompounds and, if desired, one or more additional pharmacologicallyactive substances, is regarded as coming within the protected scope ofthe present invention.

The invention also relates to processes for preparing the compound ofthe formula I.

Process A:

Process A for preparing the compound of the formula I is characterizedin that the amine of the formula II (see WO 02/50027) is, in a peptidecoupling reaction, reacted with the monoglucamide of1,12-dodecanedicarboxylic acid (formula III where the hydroxyl functionsof the glucamine moiety may be protected, for example by acyl groupssuch as acetyl groups or by ether groups such as benzyl ether groups, togive a compound of the formula IV. This reaction can be carried outusing, for example, N-hydroxy-benzotriazole (HOBt) andN-ethyl-N′-(3-dimethylaminopropyl)carbodiimide (EDC) at roomtemperature, using, for example, as solvent dimethylformamide (DMF). Itis also possible to use other peptide coupling reagents and solvents orsolvent mixtures (seer for example, A. Speicher et al. in Journal fürPraktische Chemie/Chemiker-Zeitung (1998), 340, 581-583; Y. S. Klausnerand M. Bodansky, Synthesis, (1972), 453 ff; K. Ishihara et al., J. Org.Chem., 61, 4196 (1996); M. Kunishima et al., Tetrahedron 55, 13159-13170(1999) or else R. C. Larock: Comprehensive Organic Transformations; VCH,New York, 1989, page 981 ff).

Process B:

A further process (B) according to the invention comprises reacting theamine of the formula II with 1,12-dodecanedicarboxylic acid V underpeptide coupling conditions and further reacting the product of theformula VI with glucamine VII, whose hydroxyl functions may carryprotective groups (for example acetyl protective groups or benzylprotective groups), again under peptide coupling conditions, to give thecompound of the formula I or the corresponding compound Ia which carriesprotective groups. In a further step, the protective groups may beremoved either under weakly alkaline conditions, for example diluteaqueous ammonia, or hydrogenolytically (in the case of benzyl etherprotective groups being used) to give the compound of the formula I.

Process C:

In a further process C according to the invention, the amine of theformula II is reacted with an acid halide, for example the chloride of11-((4R,6R)-4,5,6-trihydroxy-3-(R)-hydroxy-2-(S)-hydroxyhexylcarbamoyl)-undecanoicacid VIII, for example in pyridine or in dichloromethane, with orwithout addition of amine base, at room temperature. Here, the hydroxylfunctions of the glucamine moiety advantageously carry theabovementioned protective groups which may, after coupling to give theamide of the formula Ia, be removed.

The invention furthermore relates to the intermediates of formulae III,IV and VIII wherein R is acyl, for example acetyl or benzoyl, or inwhich R is aralkyl, alkyl or aryl, for example benzyl.

The citation of any reference herein should not be construed as anadmission that such reference is available as “Prior Art” to the instantapplication.

The present invention is not to be limited in scope by the specificembodiments describe herein. Indeed, various modifications of theinvention in addition to those described herein will become apparent tothose skilled in the art from the foregoing description and theaccompanying figures. Such modifications are intended to fall within thescope of the appended claims.

Various publications are cited herein, the disclosures of which areincorporated by reference in their entireties.

EXAMPLE I Process A1 1.) Monomethyl Dodecanedioate

With heating, 4.6 g (20 mmol) of dodecanoic acid are dissolved in 40 mlof dry THF, 0.73 ml (10 mmol) of thionyl chloride is added slowly andthe mixture is stirred at RT for 30 min. 0.8 ml (20 mmol) of drymethanol is then added slowly and the mixture is stirred at RT for 4 h;the mixture is then allowed to stand at RT for 4 days. After this periodof time, TLC shows no further conversion; the reaction mixture isconcentrated under reduced pressure and the residue is triturated withwater (ultrasonic bath). The precipitate is filtered off with suction,washed with water and again filtered off with suction. The moist residueis triturated with dichloromethane (ultrasonic bath), filtered through apleated filter and washed with dichloromethane, and the filtrate isconcentrated under reduced pressure. This gives monomethyldodecanedioate (3.09 g) in a yield of 63%. MW: 244.34; MS: 245.4 (M+H⁺).

2.) Synthesis of methyl11-((4R,6R)-4,5,6-trihydroxy-3-(R)-hydroxy-2-(S)-hydroxyhexylcarbamoyl)undecanoate

At room temperature, 3.07 g (12.6 mmol) of monomethyl dodecane-dioateare dissolved in 30 ml of dry DMF, 2.2 g (12.1 mmol) of glucamine, 1.9 g(12.4 mmol) of HOBt and 2.4 g (12.5 mmol) of EDC are added and themixture is stirred at RT for 6 h. The mixture is allowed to stand at RTovernight. The next day, TLC shows complete conversion. The reactionmixture is concentrated under reduced pressure and dried under highvacuum. The residue is triturated with water (ultrasonic bath), filteredoff with suction, washed with water and filtered off with suction. Themoist crude product is triturated with dichloromethane, filtered offwith suction, washed with dichloromethane and dried. This gives methyl11-((4R,6R)-4,5,6-trihydroxy-3-(R)-hydroxy-2-(S)-hydroxyhexyl-carbamoyl)undecanoate.4.45 g (90% yield). MW: 407.51; MS: 408.20 (M+H⁺).

3.) Synthesis of11-((4R,6R)-4,5,6-trihydroxy-3-(R)-hydroxy-2-(S)-hydroxyhexylcarbamoyl)undecanoicacid (III; R=H)

At room temperature, 4.45 g (10.9 mmol) of methyl11-((4R,6R)-4,5,6-trihydroxy-3-(R)-hydroxy-2-(S)-hydroxyhexyl-carbamoyl)undecanoateare suspended in 75 ml of dry ethanol, and 25 ml of water and 2.2 g ofKOH (85% strength) (33 mmol) are added. After 2 h of stirring at 80° C.,TLC shows complete conversion. The reaction mixture is concentratedunder reduced pressure; the residue is dissolved in water and acidifiedwith conc. hydrochloric acid. The precipitated crude product is filteredoff with suction, washed with water and filtered off with suction. Themoist crude product is recrystallized from about 100 ml of ethanol,filtered while hot and precipitated in an ice bath. The precipitate isfiltered off with suction, washed with ethanol and dried. This gives 2.2g (51%) of11-((4R,6R)-4,5,6-trihydroxy-3-(R)-hydroxy-2-(S)-hydroxyhexyl-carbamoyl)undecanoicacid. MW: 393.48; MS: 394.28 (M+H⁺).

4.) Synthesis of dodecanedioic acid4-[(2S,3R)-3-[(S)-3-(4-fluorophenyl)-3-hydroxypropyl]-2-(4-methoxyphenyl)-4-oxoazetidin-1-yl]benzylamide((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amide (I)

With gentle heating, 0.63 g (1.45 mmol) of benzylamine II (preparationsee DE 10064398) and 0.65 g (1.65 mmol) of the diacid monoamide (seeabove) are dissolved in 15 ml of dry DMF, 0.25 g (1.63 mmol) of HOBt and0.31 g (1.67 mmol) of EDC are added and the mixture is stirred at RT for4 h. The reaction mixture is allowed to stand at RT overnight. The nextmorning, TLC shows complete conversion. The reaction mixture isconcentrated under reduced pressure and the residue is dried under highvacuum. The residue is triturated with water (ultrasonic bath), filteredoff with suction, washed with water and filtered off with suction. Thecrude product is recrystallized from isopropanol. The crystals arefinally triturated with water, filtered off with suction and dried. Thisgives 0.38 g (32%) of dodecanedioic acid4-[(2S,3R)-3-[(S)-3-(4-fluorophenyl)-3-hydroxy-propyl]-2-(4-methoxyphenyl)-4-oxoazetidin-1-yl]benzylamide((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amide (I). MW: 809.97; MS:810.49 (M+H⁺).

Process A2 1.)11-((2S,3R,4R,5R)-2,3,4,5,6-pentaacetoxyhexylcarbamoyl)-undecanoic acid(III; R=acetyl)

At room temperature, 3 ml of dry pyridine and 3 ml of acetic anhydrideare added to 0.4 g of11-((4R,6R)-4,5,6-trihydroxy-3-(R)-hydroxy-2-(S)-hydroxyhexylcarbamoyl)undecanoicacid (III; R=H), and the mixture is stirred at room temperature for 4 h.After the reaction has ended, water is added to the reaction mixture andthe mixture is concentrated under reduced pressure. The residue istriturated with a little water and filtered. The filter residue iswashed with water and then dried under reduced pressure. This gives 0.56g of 11-((2S,3R,4R,5R)-2,3,4,5,6-pentaacetoxyhexylcarbamoyl)-undecanoicacid. MW: 603.66; MS: 604.22 (M+H⁺).

2.)(2R,3R,4R,5S)-2,3,4,5-tetraacetoxy-6-(11-{4-[(2S,3R)-3-[(S)-3-(4-fluorophenyl)-3-hydroxypropyl]-2-(4-methoxyphenyl)-4-oxoazetidin-1-yl]benzylcarbamoyl}undecanoylamino)hexylacetate (IV; R=acetyl)

At room temperature, 87 mg of amine II are dissolved in 3 ml of drieddimethylformamide, and 120 mg of the carboxylic acid described above, 31mg of N-hydroxybenzotriazole and 39 mg ofN-ethyl-N′-(3-dimethylaminopropyl)carbodiimide are added. The reactionmixture is stirred at room temperature overnight and then concentratedunder reduced pressure The residue is taken up in ethyl acetate and theorganic phase is washed with water and dried over magnesium sulfate. Themixture is then filtered and the filtrate is concentrated under reducedpressure. This gives 90 mg of(2R,3R,4R,5S)-2,3,4,5-tetraacetoxy-6-(11-{4-[(2S,3R)-3-[(S)-3-(4-fluorophenyl)-3-hydroxypropyl]-2-(4-methoxyphenyl)-4-oxoazetidin-1-yl]benzylcarbamoyl}undecanoylamino)hexylacetate. MW: 1020.16.

3.) Dodecanedioic acid4-[(2S,3R)-3-[(S)-3-(4-fluorophenyl)-3-hydroxy-propyl]-2-(4-methoxyphenyl)-4-oxoazetidin-1-yl]benzylamide((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amide (I)

90 mg of the compound described above are treated with guanidine in amixture of ethanol and dichloromethane. This gives the glucaminederivative I of MW 809.97.

Process B 1.)11-{4-[(2S,3R)-3-[(S)-3-(4-fluorophenyl)-3-hydroxypropyl]-2-(4-methoxyphenyl)-4-oxoazetidin-1-yl]benzylcarbamoyl}undecanoicacid (VI)

A solution of 70 mg of amine I and 23 μl of triethylamine in 1 ml ofdimethylformamide is added to a solution of 371 mg of dodecanedioicacid, 63 μl of diisopropylcarbodiimide and 55 mg of hydroxybenzotriazolein 2 ml of dimethylformamide, and the mixture is stirred at roomtemperature for 12 h. The reaction solution is concentrated andseparated by HPLC (Knauer Eurospher-100-10-C18, water (0.1%trifluoroacetic acid)/acetonitrile (0.1% trifluoroaceticacid)=80/20→10/90). This gives the product of molecular weight 646.81(C₃₈H₄₇F₁N₂O₆); MS (ESI) 647.35 (M+H⁺)

2.) Dodecanedioic acid4-[(2S,3R)-3-[(S)-3-(4-fluorophenyl)-3-hydroxypropyl]-2-(4-methoxyphenyl)-4-oxoazetidin-1-yl]benzyl-amide((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amide (I)

As described above for other coupling reactions, reaction of acid VIwith glucamine (VII; R=H) and HOBt/EDC in DMF gives the compound I(R=H).

If, instead of glucamine, a protected glucamine derivative, for exampleVII (R=acetyl) is used, the compound Ia where R=acetyl is obtained.

Process C 1.)(2R,3R,4R,5R)-2,3,4,5-tetraacetoxy-6-(11-chlorocarbonyl-undecanoylamino)hexylacetate (VIII; R=acetyl)

The compound of the formula III (R=acetyl) is dissolved intetrahydrofuran and thionyl chloride is added slowly; the mixture isstirred at room temperature for 1 h. The reaction solution is thenconcentrated under reduced pressure and the crude product is used forthe next step.

2.)(2R,3R,4R,5S)-2,3,4,5-tetraacetoxy-6-(11-{4-[(2S,3R)-3-[(S)-3-(4-flurophenyl)-3-hydroxypropyl]-2-(4-methoxyphenyl)-4-oxoazetidin-1-yl]benzylcarbamoyl}undecanoylamino)hexylacetate (IV; R=acetyl)

At room temperature, amine I is added to the acid chloride shown abovein a mixture of pyridine and dichloromethane, and the mixture is stirredat room temperature overnight. Work-up gives amide IV where R=acetyl.

Using the method described below, the activity of the compound of theformula I according to the invention was examined:

Effect on Cholesterol Absorption+³H-Taurocholic Acid Excretion UsingFecal Excrement of Mice, Rats or Hamsters

NMRI mice, Wistar rats, or Golden Syrian hamsters (in groups of n=4-6)are kept in metabolic cages, where they are fed with a standard diet(Altromin, Lage (Lippe)). The afternoon prior to the administration ofthe radioactive tracers (¹⁴C-cholesterol), the feed is removed and theanimals are adapted to grates.

Additionally, the animals are labeled s.c with ³H-TCA (taurocholic acid)(for example 1 μCi/mouse up to 5 μCi/rat) 24 hours prior to the peroraladministration of the test meal (¹⁴C-cholesterol in Intralipid® 20,Pharmacia-Upjohn).

Cholesterol absorption test: 0.25 ml/mouse Intralipid® 20(Pharmacia-Upjohn) ((spiked with 0.25 μCi of ¹⁴C-cholesterol in 0.1 mgof cholesterol) is administered perorally by gavage.

Test substances are prepared separately in 0.5%/(methylcellulose(Sigma)/5% Solutol (BASF, Ludwigshafen) or a suitable vehicle.

The administration volume of the test substance is 0.5 ml/mouse. Thetest substance is administered immediately prior to the test meal(Intralipid labeled with ¹⁴C-cholesterol) (cholesterol absorption test).

The feces are collected over a period of 24 h: fecal elimination of¹⁴C-cholesterol and ³H-taurocholic acid (TCA) is determined after 24hours.

The livers are removed and homogenized, and aliquots are incinerated inan oximate (Model 307, Packard) to determine the amount of¹⁴C-cholesterol which had been taken up/absorbed.

Evaluation:

Feces Samples:

The total weight is determined, the sample is made up with water to adefined volume and then homogenized, and an aliquot is evaporated todryness and incinerated in an oximate (Model 307 from Packard for theincineration of radioactively labeled samples): the amount ofradioactive ³H—H2O and ¹⁴C—CO2 is extrapolated to the amount of³H-taurocholic acid and ¹⁴C-cholesterol, respectively, that is excreted(dual isotope technique). The ED₂₀₀ values as dose from a dose-effectcurve are interpolated as those doses at which the excretion of TCA orcholesterol is doubled, based on a control group treated at the sametime.

Liver Samples:

The amount of ¹⁴C-cholesterol taken up by the liver is based on theadministered dose. The ED₅₀ values are interpolated from a dose-effectcurve as the dose at which the uptake of ¹⁴C-cholesterol by the liver ishalved (50%), based on a control group.

The ED₅₀ value below demonstrates the activity of the compound of theformula I according to the invention

Example No. ED₅₀ (liver) [mg/mouse] I 0.005

As can be seen from the table, the compound of the formula I has verygood cholesterol-lowering action.

The solubility of compound I and of the comparative compound C1 wastested as follows:

The comparative compound selected was the compound from WO 02/50027 withthe most similar structure:

0.5 mg of the compound to be tested was weighed exactly into anEppendorf cap, and 0.5 ml of the solvent in question (aqueous buffer)was added. The Eppendorf cap was then introduced into a thermomixer and,at 25° C., shaken at 1 400 rpm for 4 hours.

The Eppendorf cap was then introduced into a centrifuge. Followingcentrifugation, an aliquot of the supernatant was used to determine theamount dissolved, using HPLC/UV analysis. The table below shows theresults obtained:

Example 1 C1 pH conditions Solubility in μg/ml Solubility in μg/ml Water(pH) 3 (6.8) <1 pH 1.2 3 <1 pH 4.5 4 <1 pH 6.8 2 <1 pH 8.0 2 <1 FaSSIF28 5 FeSSIF 454 18

In the physiological solvents FaSSIF and FeSSIF (composition andpreparations see Physiologically based dissolution tests—Experienceswith poorly soluble drugs, January 2000, Shaker, ISBN: 3-8265-6962-8)the solubility of example 1 was determined as being 28 and 454 μg/ml,respectively, whereas the corresponding values for C1 were 5 and 18μg/mi, respectively. This significantly different solubility could alsobe confirmed during a repetition of the tests (43/290 μg/ml compared to6/20 μg/ml).

Accordingly, the compound of the formula I according to the inventionhas a 6-fold to 16-fold better solubility than the comparative compoundof the formula C1. Accordingly, the compound of the formula I accordingto the invention has better availability in dissolved form at the siteof action. In contrast to the more poorly soluble substances, even anyhigher doses can be provided completely for interaction with thetransport system in question. Based on an available volume of 250 ml(Biopharmaceutical Classification System), doses of up to ˜100 mg aresoluble, whereas in the case of C1 in the best case only doses in therange of 5 mg would be soluble (in the worst case even only: 1.25 mg).

The stability of compound 1 and that of the comparative compound C1 insolution was tested as follows:

The stability of dissolved compound I and of dissolved C1 was determinedin aqueous buffers in the pH range 1.2-8.0. 1 mg of the compound inquestion was weighed into a 5 ml measuring flask. A small amount ofacetonitrile was used to dissolve the substance. The flask was thenfilled up to the mark with the aqueous buffer. The precipitated compoundwas centrifuged and the clear supernatant was then tested for stabilityin solution for 24 hours at 37° C. The samples were evaluated usingHPLC/UV. The results obtained for example I and C1 are shown in thetable below:

Example I C1 Increase of the area of Increase of the area of the pHconditions the impurities in percent impurities in percent pH 1.2 4.913.3 pH 6.8 0 0.5 pH 8.0 0.2 4.6

Thus, depending on the pH, the compound of the formula I according tothe invention is at least 2.7 times more stable than C1 and,accordingly, forms fewer by-products than C1. Smaller amounts ofby-products with systemic action mean a reduced potential for unwantedside-effects.

1. A compound of formula I